Pratylenchus speijeri n. sp. (Nematoda: Pratylenchidae), a new root-lesion nematode pest of plantain in West Africa

Nematology ◽  
2012 ◽  
Vol 14 (8) ◽  
pp. 987-1004 ◽  
Author(s):  
Francesca De Luca ◽  
Alberto Troccoli ◽  
Larry W. Duncan ◽  
Sergei A. Subbotin ◽  
Lieven Waeyenberge ◽  
...  
Keyword(s):  
New Species ◽  
West Africa ◽  
Species Complex ◽  
Rrna Gene ◽  
28S Rrna ◽  
New Taxon ◽  
Root Lesion Nematode ◽  
Specific Primers ◽  
Lesion Nematode ◽  
Species Specific

A new root-lesion nematode, particularly pathogenic to Musa spp. and causing important plantain losses in Ghana, is described and named Pratylenchus speijeri n. sp. The cryptic status of this species within the P. coffeae species complex has been assessed and confirmed in this study. An extensive comparison of the morphological and molecular characteristics of this new species with those of P. coffeae and other related amphimictic species did not result in an unambiguous separation of this species from P. coffeae because only a few morphological features of diagnostic value were found. Sequence and phylogenetic analyses of the D2-D3 expansion segments of the 28S rRNA gene, the ITS rRNA gene and a portion of the hsp90 gene of P. speijeri n. sp. and P. coffeae species complex populations from different sources generated majority consensus BI trees with three major clades: P. speijeri n. sp. from Musa spp. roots in Ghana; unidentified or putative new Pratylenchus sp. C1 from Colocasia esculenta roots in Japan and P. coffeae with non-homogeneous relationships from different hosts and distant geographical areas. These results confirmed the validity of P. speijeri n. sp. as a new taxon and indicated that P. coffeae populations from Colocasia in Japan also need to be considered as a new species. Sequence differences in the ITS were used to design group- and species-specific primers to detect P. speijeri n. sp. and other species of P. coffeae species complex. The use of these species-specific primers for the separation of P. speijeri n. sp., Pratylenchus sp. C1 and P. coffeae has important practical application in breeding programmes for agriculture in West Africa.

Description of Tylenchulus musicola sp. n. (Nematoda: Tylenchulidae) from banana in Iran with molecular phylogeny and characterisation of species of Tylenchulus Cobb, 1913

Nematology ◽  
2012 ◽  
Vol 14 (3) ◽  
pp. 353-369 ◽  
Author(s):  
Zahra Tanha Maafi ◽  
Majid Amani ◽  
Jason D. Stanley ◽  
Renato N. Inserra ◽  
Esther Van den Berg ◽  
...  
Keyword(s):  
New Species ◽  
Rapid Identification ◽  
Rrna Genes ◽  
Rrna Gene ◽  
28S Rrna ◽  
Specific Primers ◽  
Diagnostic Pcr ◽  
Second Stage ◽  
Species Specific ◽  
A New Species

During a survey conducted on banana plantations in Sistan and Blouchestan province, south-east Iran, a new species of Tylenchulus was extracted from the soil and roots of banana plants. This species, named Tylenchulus musicola sp. n., is characterised by mature females having a swollen, hook-shaped body with a conical and elongate post-vulval portion ending in a round terminus, males having a weak stylet and a cylindrical and thick tail ending in a bluntly rounded and smooth terminus, and by second-stage juveniles having a slender body and a posterior body portion ending in a finely pointed or mucronate terminus. The results of glasshouse host tests indicated that the new species does not parasitise sugarcane ratoons or sour orange seedlings. Tylenchulus musicola sp. n. is distinguished from other known Tylenchulus species by the sequences of D2-D3 expansion segments of 28S rRNA and ITS rRNA genes. Phylogenetic relationships within Tylenchulus were reconstructed based on rRNA gene sequences using Bayesian inference. Diagnostic PCR-ITS-RFLP profiles are presented for T. musicola sp. n., T. furcus, T. graminis, T. palustris, T. semipenetrans and Trophotylenchulus floridensis. PCR with species-specific primers and genus-specific primer are tested and developed for rapid identification of five Tylenchulus species. An identification key to Tylenchulus species is provided.

Ditylenchus gallaeformans sp. n. (Tylenchida: Anguinidae) – a neotropical nematode with biocontrol potential against weedy Melastomataceae

Nematology ◽  
2013 ◽  
Vol 15 (2) ◽  
pp. 179-196 ◽  
Author(s):  
Rosângela D.L. Oliveira ◽  
Ângelo M. Santin ◽  
Dalila J. Seni ◽  
Anna Dietrich ◽  
Luis A. Salazar ◽  
...  
Keyword(s):  
New Species ◽  
Severe Disease ◽  
Molecular Study ◽  
Rrna Gene ◽  
28S Rrna ◽  
Leaf Spots ◽  
Miconia Calvescens ◽  
Biocontrol Potential ◽  
Tail Tip ◽  
Species Specific

Ditylenchus gallaeformans sp. n. was found on several hosts at numerous locations in Brazil and Costa Rica. In its native habitats it attacks several genera in the Melastomataceae, including two species ranked as among the worst invasive weeds of Pacific island forests, namely Miconia calvescens and Clidemia hirta. The new species causes a severe disease on infected plants involving the formation of gall-like structures on infected leaves, inflorescences and stems, and may cause significant impact on its hosts. Morphological study using light and scanning electron microscopy and analysis of the partial 18S rRNA, the D2-D3 expansion fragments of 28S rRNA and the ITS rRNA gene sequences showed little variations between populations from different hosts or geographical origins. The molecular study revealed that the new species is related to D. drepanocercus, which was recently found in association with M. calvescens but causing angular leaf spots on this host. Ditylenchus gallaeformans sp. n. is distinguished from D. drepanocercus by having a bursa reaching the tail tip (vs covering around 50% of tail in D. drepanocercus) and a conical tail, regularly tapering towards a variable tip (vs tail with a distinctive apical falciform appendage in both sexes in D. drepanocercus). PCR with species-specific primers was developed for diagnostics of both Ditylenchus species. Ditylenchus gallaeformans sp. n. deserves further investigation as a potential biocontrol agent against M. calvescens and C. hirta.

scholarly journals Pratylenchus vovlasi sp. Nov. (Nematoda: Pratylenchidae) on Raspberries in North Italy with a Morphometrical and Molecular Characterization

Plants ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 1068
Author(s):  
Alberto Troccoli ◽  
Elena Fanelli ◽  
Pablo Castillo ◽  
Gracia Liébanas ◽  
Alba Cotroneo ◽  
...  
Keyword(s):  
Its Region ◽  
Nematode Species ◽  
Integrative Taxonomy ◽  
Integrative Approach ◽  
Parasitic Nematodes ◽  
Rrna Gene ◽  
28S Rrna ◽  
Root Lesion Nematode ◽  
Cyst Nematodes ◽  
Lesion Nematode

Root-lesion nematode species rank third only to root-knot and cyst nematodes as having the greatest economic impact on crops worldwide. A survey of plant-parasitic nematodes associated with decaying raspberries (Rubus sp.) in northern Italy revealed that root-lesion nematodes were the most frequently occurring species among other phytonematodes. Several Pratylenchus species have been associated with Rubus sp. in Canada (Quebec, British Columbia) and USA (North Carolina, Maryland, New Jersey) including P. penetrans and P. crenatus. In the roots and rhizosphere of symptomatic raspberries, nematodes of two Pratylenchus spp. were detected. Detailed morphometrics of the two root-lesion nematode isolates were consistent with Pratylenchus crenatus and with an undescribed Pratylenchus species. The extracted nematodes were observed and measured as live and fixed materials and subsequently identified by integrative taxonomy (morphometrically and molecularly). The latter species is described herein as Pratylenchus vovlasi sp. nov., resulting morphometrically closest to P. mediterraneus and phylogenetically to P. pratensis. The molecular identification of Pratylenchus vovlasi sp. nov. was carried out by sequencing the ITS region, D2-D3 expansion domains of the 28S rRNA gene and a partial region of the nuclear hsp90 gene. ITS-RFLP and sequence analyses revealed that Pratylenchus vovlasi sp. nov. had species-specific restriction profiles with no corresponding sequences present in the database. The phylogenetic relationships with ITS and D2-D3 sequences placed the Pratylenchus vovlasi sp. nov. in a clade with P. pratensis and P. pseudopratensis. This research confirms the occurrence of cryptic biodiversity within the genus Pratylenchus as well as the need for an integrative approach to the identification of Pratylenchus species.

scholarly journals Discovery of Root-Lesion Nematode (Pratylenchus scribneri) on Corn in Hainan Province of China

Plant Disease ◽  
2021 ◽  
Author(s):  
Yanhui Xia ◽  
Jing Li ◽  
Penghui Hao ◽  
Ke Wang ◽  
Bin Lei ◽  
...  
Keyword(s):  
Its Sequences ◽  
Rrna Gene ◽  
Hainan Province ◽  
28S Rrna ◽  
Root Lesion Nematode ◽  
28S Rrna Gene ◽  
Mtdna Coi ◽  
Lesion Nematode ◽  
Corn Plants ◽  
Pratylenchus Scribneri

Corn (Zea mays L.) is a very important cereal crop and serves as food, feed, and industrial material (Liu et al. 2016). The root-lesion nematode (RLN) is considered one of the most important plant-parasitic nematodes and can cause economic losses in agriculture worldwide (Jones et al. 2013). In January 2020, five samples were collected from a corn field in Lingshui Lizu Autonomous County, Hainan Province, China. The collected corn plants (cv. Denghai 685) were growing poorly and roots showed distinct lesions and rot. Corn fields with symptoms of stunted plants, and brown lesions on roots were widespread. This corn disease was severe in Lingshui Lizu Autonomous County. RLN were extracted from soil samples by the modified Baermann funnel (Hooper et al. 2005). All the samples contained RLN ranging from 9 to 82 (average 39) RLN per 100 cm3 of soil and 113 to 257 (average 194) RLN per 5 g roots. The extracted RLN were sterilized and cultured on carrot disks at 25°C for 90 days. Afterwards, seeds of corn (cv. Denghai 685) were sown in pots containing 1.8 liters of sterilized soil. Eight plants, one per pot, were infested with 1,000 RLN, eight pots of noninfested corn plants were used as controls, and plants were kept in a greenhouse at 25°C. At 75 days after inoculation, symptoms were like those initially observed in corn fields, whereas no symptoms were observed in the control plants. Nematodes in the soil and roots were extracted using the same method as previously described (Hooper et al. 2005). The average number of RLN per pot was approximately 4,250 in soil and 820 in roots, the reproduction factor (final number of nematodes/initial number of nematodes) was 5.07, no RLN were found in the control. The experiment was conducted twice. The morphological and molecular studies of RLN were examined to confirm species identification. The main morphological measurements of adult females (n = 15) included body length = 526.0 μm ± 17.1 (standard error) (range = 498.0 to 560.5 μm), stylet = 16.0 μm ± 0.3 (15.5 to 16.5 μm), tail length = 29.0 μm ± 1.5 (26.5 to 31.0 μm), a = 23.6 ± 0.6 (22.6 to 24.4), b = 5.6 ± 0.3 (5.2 to 6.0), c = 18.3 ± 0.9 (16.4 to 19.7), V = 78.2% ± 0.6 (77.4 to 79.2%), lip region with two annules. No males were found in the samples. This population was identified as Pratylenchus scribneri, based on the morphological characters (Castillo and Vovlas, 2007). DNA was isolated from individual nematodes followed by proteinase K-based lysis (Wang et al. 2011). The D2/D3 expansion region of the 28S rRNA gene, rDNA-internal transcribed spacer (ITS) region, and mitochondrial cytochrome oxidase I (mtDNA-COI) gene were amplified with primers D2A/-D3B (De Ley et al. 1999), TW81/ AB28 (Vovlas et al. 2011) and JB3/ JB5 (Liu et al. 2018), respectively. The PCR products were purified and ligated into pJET 1.2/blunt cloning vectors and transformed to Escherichia coli strain DH5α, and then sequenced. The obtained 28S rRNA gene D2/D3 region sequences (785bp), ITS sequences (886 bp) and mtDNA-COI (447bp) in this study were submitted to GenBank. The D2/D3 region of the 28S rRNA sequences of the RLN collected in Lingshui (GenBank accession no. MZ701843) showed 99.75% identity with P. scribneri sequences available in the GenBank (KX842628 and KX842625). The ITS sequences of the RLN collected in this study (MZ701842) showed the highest identity of 97.06% with P. scribneri sequences available in the GenBank (KX842626). The mtDNA-COI sequences of the RLN collected in this study (OK040228) showed 100% identity with P. scribneri (MN366409). Both morphological and molecular data confirmed the identity of P. scribneri. P. scribneri has been reported on corn in Inner Mongolia, Hebei, Shanxi, Shandong, Henan, Jiangsu, and Liaoning provinces of China (Li et al. 2019). As far as we know, this is the first report of P. scribneri on corn in Hainan Province, China. Since the RLN can cause considerable damage to corn, strategic measures should be taken to prevent the spread of P. scribneri to other regions in China.

Morphological and molecular characterisation of some Hemicriconemoides species (Nematoda: Criconematidae) together with a phylogeny of the genus

Nematology ◽  
2014 ◽  
Vol 16 (5) ◽  
pp. 519-553 ◽  
Author(s):  
Esther Van den Berg ◽  
Louwrens R. Tiedt ◽  
Renato N. Inserra ◽  
Jason D. Stanley ◽  
Nicola Vovlas ◽  
...  
Keyword(s):  
Fruit Trees ◽  
Valid Species ◽  
Rrna Gene ◽  
28S Rrna ◽  
Molecular Characterisation ◽  
Electron Microscopic ◽  
Specific Primers ◽  
Scanning Electron Microscopic ◽  
The Usa ◽  
Species Specific

Sheathoid nematodes of the genusHemicriconemoidesare migratory root-ectoparasites of many plants including various agricultural crops and fruit trees. They are generally found inhabiting warm areas of the world and presently consist of 52 valid species. In this study we provide morphological and molecular characterisation of 12 species of this genusviz.:H. alexis,H. brachyurus,H. californianus,H. chitwoodi,H. macrodorus,H. minutus,H. ortonwilliamsi,H. promissus,H. silvaticus,H. strictathecatus,H. wessoniandHemicriconemoidessp. originating from China, Greece, Japan, Myanmar, Spain, South Africa and the USA. Morphological descriptions, measurements, light and scanning electron microscopic observations and drawings are given for several species. Phylogenetic relationships withinHemicriconemoides, as inferred from the analyses of the D2-D3 of 28S rRNA and ITS-rRNA gene sequences, resulted in trees with three major clades that corresponded with species groupings based on morphology of the lip pattern and vulval flap. PCR with species-specific primers were developed forH. californianus,H. chitwoodiandH. strictathecatus.

Description of a new species of Auriculostoma (Digenea: Allocreadiidae) from Characidium heirmostigmata (Characiformes: Crenuchidae) from Argentina, using morphological and molecular data

2021 ◽  
Vol 95 ◽  
Author(s):  
M.M. Montes ◽  
J. Barneche ◽  
Y. Croci ◽  
D. Balcazar ◽  
A. Almirón ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
New Species ◽  
National Park ◽  
Molecular Data ◽  
Rrna Gene ◽  
28S Rrna ◽  
The Family ◽  
Parasitological Survey ◽  
Morphological And Molecular Data ◽  
A New Species

Abstract During a parasitological survey of fishes at Iguazu National Park, Argentina, specimens belonging to the allocreadiid genus Auriculostoma were collected from the intestine of Characidium heirmostigmata. The erection of the new species is based on a unique combination of morphological traits as well as on phylogenetic analysis. Auriculostoma guacurarii n. sp. resembles four congeneric species – Auriculostoma diagonale, Auriculostoma platense, Auriculostoma tica and Auriculostoma totonacapanensis – in having smooth and oblique testes, but can be distinguished by a combination of several morphological features, hosts association and geographic distribution. Morphologically, the new species can be distinguished from both A. diagonale and A. platense by the egg size (bigger in the first and smaller in the last); from A. tica by a shorter body length, the genital pore position and the extension of the caeca; and from A. totonacapanensis by the size of the oral and ventral sucker and the post-testicular space. Additionally, one specimen of Auriculostoma cf. stenopteri from the characid Charax stenopterus (Characiformes) from La Plata River, Argentina, was sampled and the partial 28S rRNA gene was sequenced. The phylogenetic analysis revealed that A. guacurarii n. sp. clustered with A. tica and these two as sister taxa to A. cf. stenopteri. The new species described herein is the tenth species in the genus and the first one parasitizing a member of the family Crenuchidae.

Species-specific duplex PCR for the detection of Pratylenchus penetrans

Nematology ◽  
2009 ◽  
Vol 11 (6) ◽  
pp. 847-857 ◽  
Author(s):  
Lieven Waeyenberge ◽  
Nicole Viaene ◽  
Maurice Moens
Keyword(s):  
Internal Control ◽  
Dna Sequences ◽  
Rrna Gene ◽  
Duplex Pcr ◽  
Specific Primers ◽  
5.8S Rrna Gene ◽  
5.8S Rrna ◽  
Wide Range ◽  
Pcr Products ◽  
Species Specific

Abstract ITS1, the 5.8S rRNA gene and ITS2 of the rDNA region were sequenced from 20 different Pratylenchus species. Additionally, the same region was sequenced from seven populations of P. penetrans. After purifying, cloning and sequencing the PCR products, all sequences were aligned in order to find unique sites suitable for the design of species-specific primers for P. penetrans. Since ITS regions showed variability between and even within populations of P. penetrans, only three small DNA sequences were suitable for the construction of three potentially useful species-specific primers. New species-specific primers were paired with existing universal ITS primers and tested in all possible primer combinations. The best performing primer set, supplemented with a universal 28S rDNA primer set that served as an internal control, was tested in duplex PCR. The ideal annealing temperature, Mg2+ concentration and primer ratios were then determined for the most promising primer set. The optimised duplex PCR was subsequently tested on a wide range of different Pratylenchus spp. and 25 P. penetrans populations originating from all over the world. To test the sensitivity, the duplex PCR was conducted on DNA extracted from a single P. penetrans nematode mixed with varying amounts of nematodes belonging to another Pratylenchus species. Results showed that a reliable and sensitive P. penetrans species-specific duplex PCR was constructed.

scholarly journals Multiplex PCR with 16S rRNA Gene-Targeted Primers of Bifidobacterium spp. To Identify Sources of Fecal Pollution

2004 ◽  
Vol 70 (5) ◽  
pp. 3171-3175 ◽  
Author(s):  
X. Bonjoch ◽  
E. Ballesté ◽  
A. R. Blanch
Keyword(s):  
16S Rrna ◽  
Multiplex Pcr ◽  
Fecal Pollution ◽  
Sensitivity Threshold ◽  
Rrna Gene ◽  
Specific Primers ◽  
Animal Wastewater ◽  
Wastewater Samples ◽  
Species Specific ◽  
Different Origins

ABSTRACT Bifidobacteria are one of the most common bacterial types found in the intestines of humans and other animals and may be used as indicators of human fecal pollution. The presence of nine human-related Bifidobacterium species was analyzed in human and animal wastewater samples of different origins by using species-specific primers based on 16S rRNA sequences. Only B. adolescentis and B. dentium were found exclusively in human sewage. A multiplex PCR approach with strain-specific primers was developed. The method showed a sensitivity threshold of 10 cells/ml. This new molecular method could provide useful information for the characterization of fecal pollution sources.

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